TABLE 1.
Bacterial strains used in this study
Antibiotics were used at the following concentrations: for E. coli cultures: kanamycin (KanR) 30 μg/ml, ampicillin (AmpR) 100 μg/ml, gentamicin (GnR) 20 μg/ml, and chloramphenicol (CamR) 10 μg/ml; for S. aureus cultures: erythromycin (ErmR) 10 μg/ml, kanamycin (KanR) 90 μg/ml, and chloramphenicol (CamR) 7.5 μg/ml. Inducers were used at the following concentrations: isopropyl 1-thio-β-d-galactopyranoside 1 mm; Atet 50 ng/ml. nt means nucleotide.
| Strain | Relevant features | Ref. |
|---|---|---|
| Escherichia coli strains | ||
| XL1-Blue | Cloning strain, TetR ANG127 | Stratagene |
| DH5α | Cloning strain-ANG397 | 56 |
| ANG203 | pCN49 in E. coli: AmpR | 34 |
| ANG292 | pCL55iTETr862 in XL1-Blue: AmpR | 31 |
| ANG1429 | pKOR1 in E. coli strain DB3.1: AmpR | 36 |
| ANG1632 | pCN34iTET in E. coli: AmpR | 22 |
| ANG1676 | pCN38 in XL1-Blue: AmpR | 34 |
| ANG1861 | pET28b-gdpP84–655 in BL21 (DE3): KanR | 22 |
| ANG1970 | pET28b-disA in BL21 (DE3): KanR | 22 |
| ANG1971 | pET28b-dacA in BL21 (DE3): KanR | 22 |
| ANG2246 | pCN49iTET in XL1-Blue: AmpR | This study |
| ANG2247 | pCN49iTET-dacA in XL1-Blue: AmpR | This study |
| ANG2250 | pKOR1-dacA::kan in DH5α: AmpR | This study |
| ANG2352 | pREP4 in XL1-Blue: KanR | Invitrogen |
| ANG2547 | pCL55iTETr862-dacA in XL1-Blue pREP4: AmpR, KanR | This study |
| ANG2815 | pVL847 in XL1-Blue: AmpR | 57 |
| ANG2882 | pVL847-ktrA in XL1-Blue: AmpR | This study |
| ANG2883 | pVL847-ktrA in BL21 (DE3): AmpR | This study |
| ANG2890 | pVL847 in BL21 (DE3): AmpR | This study |
| ANG3030 | pET28b-relA in XL1-Blue: KanR | This study |
| ANG3031 | pET28b-relA in BL21 (DE3): KanR | This study |
| ANG3032 | pET28b-gppA in XL1-Blue: KanR | This study |
| ANG3033 | pET28b-gppA in BL21 (DE3): KanR | This study |
| ANG3162 | pKOR1-rshsyn in DH5α: AmpR | This study |
| ANG3189 | pVL847-rsh in T7IQ: GnR, CamR | Lab strain collection |
| Staphylococcus aureus strains | ||
| SEJ1 | RN4220spa; protein A negative derivative of RN4220; ANG314 | 58 |
| ANG467 | NMΔΦ4; Newman strain with deletion of phage NM4, which allows integration of pCL55-derived plasmids | 31 |
| LAC* | Erm sensitive CA-MRSA LAC strain (AH1263); ANG1575 | 59 |
| SH1000 | Strain 8325–4 with repaired defect in rsbU | 60 |
| ANG1961 | LAC*gdpP::kan: KanR | 22 |
| ANG2253 | SEJ1 dacA::kan pCN49iTET-dacA: KanR, ErmR | This study |
| ANG2542 | LAC* pCL55iTETr862-dacA: CamR | This study |
| ANG2543 | LAC*dacA::kan pCL55iTETr862-dacA (LAC*idacA): KanR, CamR | This study |
| ANG2545 | SEJ1 pCL55iTETr862-dacA: CamR | This study |
| ANG3163 | SEJ1 pKOR1-rshsyn: CamR | This study |
| ANG3165 | LAC*rshsyn | This study |
| ANG3188 | SH1000 ΔahpC ΔkatA | 61 |
| ANG3306 | LAC* ΔgdpP rshsyn | This study |
| HG001 | NCTC8325 rsbU repaired ANG3401 | 62 |
| HG001–230 | HG001 relQsyn: deleted for nt 343–429; ANG3402 | 17 |
| HG001–229 | HG001 relPsyn: deleted for nt 450–536; ANG3403 | 17 |
| ANG3407 | HG001 ΔgdpP | This study |
| ANG3408 | HG001 ΔgdpP relPsyn | This study |
| ANG3409 | HG001 ΔgdpP relQsyn | This study |